Phenotypic and evolutionary variation in fish myofibrillar proteins

Chapter 1 General Introduction: The general introduction initially presents the major landmarks in muscle research of the last 3 millennia. The proteins of the contractile apparatus and their role in muscle contraction are described. There is then a description of how contractile proteins are known to alter through the expression of isoforms. Finally, a description of fish muscle and its fibre types is given, followed by the aims of the thesis. Chapter 2 Materials and Methods: An account is given of the materials and methods used throughout this thesis. This includes myofibril preparation and electrophoretic techniques. The techniques described are one and two dimensional polyacrylamide gel electrophoresis (PAGE), iso-electric focusing (lEF), peptide mapping, and one and two dimensional alkali-urea PAGE. The methods used to fix, stain and store the gels are then given, followed by the protocol used for Western blotting. Finally, the analysis of proteins bands is described. Chapter 3 Temperature and the plasticity of myofibrillar proteins during ontogeny in the Atlantic herring (Clupea harengus L.): Many aspects of development are influenced by temperature. The aim of this study was to investigate the effect of rearing temperature on the development and myofibrillar protein expression during ontogeny in the Atlantic herring (Clupea harengus L.). Chapter 4 The effect of body size on the myofibrillar protein composition of fast muscle fibres in the short horn sculpin (Myoxocephalus scorpius L.): The contractile properties of muscle vary with body length in fish. The aim of this study was determine if the proteins altered with body size in the short-horn sculpin (Myoxocephalus scorpius L.). Furthermore, could changes in protein expression be related to differences in contractile properties. Chapter 5 The myofibrillar proteins of Antarctic and sub-Antarctic fish: The myosins of Antarctic fish are specialised for function at low temperature. The aim of this study was to determine if the myofibrillar proteins present in Antarctic fish were highly conserved for function in this stable, low temperature environment. The variation in protein structure from myotomal fast muscles and the m. adductor profundis between five Antarctic fish species from two genera was compared with five sub-Antarctic species from four genera. The myofibrillar proteins of both the Antarctic and sub-Antarctic species showed a high degree of similarity between fish within the same genera. However, the isoforms present were considerably different between genera in both the Antarctic and sub-Antarctic species. Furthermore, the extent of the variation in protein isoforms between genera of the Antarctic fish was similar to that of the sub-Antarctic fish. This suggests that divergence in the tertiary structure of myosins from these species has occurred and that the Antarctic fish myofibrillar proteins are not highly conserved. Chapter 6 General Discussion: The major findings of the thesis are discussed in relation to the expression of myofibrillar proteins, with reference to further studies. (Abstract shortened

Percutaneous Transendocardial Delivery of Self-complementary Adeno-associated Virus 6 Achieves Global Cardiac Gene Transfer in Canines

Achieving efficient cardiac gene transfer in a large animal model has proven to be technically challenging. Previous strategies have used cardiopulmonary bypass or dual catheterization with the aid of vasodilators to deliver vectors, such as adenovirus, adeno-associated virus (AAV), or plasmid DNA. Although single-stranded AAV (ssAAV) vectors have shown the greatest promise, they suffer from delayed expression, which might be circumvented using self-complementary vectors. We sought to optimize cardiac gene transfer using a percutaneous transendocardial injection catheter to deliver adeno-associated viral vectors to the canine myocardium. Four vectors were evaluated-ssAAV9, self-complementary AAV9 (scAAV9), scAAV8, scAAV6-so that comparison could be made between single-stranded and self-complementary vectors as well as among serotypes 9, 8, and 6. We demonstrate that scAAV is superior to ssAAV and that AAV 6 is superior to the other serotypes evaluated. Biodistribution studies revealed that vector genome copies were 15-4,000 times more abundant in the heart than in any other organ for scAAV6. Percutaneous transendocardial injection of scAAV6 is a safe, effective method to achieve efficient cardiac gene transfer

The clustering of galaxies in the SDSS-III Baryon Oscillation Spectroscopic Survey: measurements of the growth of structure and expansion rate at z=0.57 from anisotropic clustering

We analyze the anisotropic clustering of massive galaxies from the Sloan Digital Sky Survey III Baryon Oscillation Spectroscopic Survey (BOSS) Data Release 9 (DR9) sample, which consists of 264,283 galaxies in the redshift range 0.43 < z < 0.7 spanning 3,275 square degrees. Both peculiar velocities and errors in the assumed redshift-distance relation ("Alcock-Paczynski effect") generate correlations between clustering amplitude and orientation with respect to the line-of-sight. Together with the sharp baryon acoustic oscillation (BAO) standard ruler, our measurements of the broadband shape of the monopole and quadrupole correlation functions simultaneously constrain the comoving angular diameter distance (2190 +/- 61 Mpc) to z=0.57, the Hubble expansion rate at z=0.57 (92.4 +/- 4.5 km/s/Mpc), and the growth rate of structure at that same redshift (d sigma8/d ln a = 0.43 +/- 0.069). Our analysis provides the best current direct determination of both DA and H in galaxy clustering data using this technique. If we further assume a LCDM expansion history, our growth constraint tightens to d sigma8/d ln a = 0.415 +/- 0.034. In combination with the cosmic microwave background, our measurements of DA, H, and growth all separately require dark energy at z > 0.57, and when combined imply \Omega_{\Lambda} = 0.74 +/- 0.016, independent of the Universe's evolution at z<0.57. In our companion paper (Samushia et al. prep), we explore further cosmological implications of these observations.Comment: 19 pages, 11 figures, submitted to MNRAS, comments welcom

Physical Examination Variables Predict Response to Conservative Treatment of Nonchronic Plantar Fasciitis: Secondary Analysis of a Randomized, Placeboâ Controlled Footwear Study

BackgroundPlantar fasciitis is a common, disabling condition, and the prognosis of conservative treatment is difficult to predict.ObjectiveTo determine whether initial clinical findings could help predict patient response to conservative treatment that primarily consisted of supportive footwear and stretching.SettingPatients were recruited and seen at 2 outpatient podiatric clinics in the Chicago, Illinois, metropolitan area.PatientsSeventyâ seven patients with nonchronic plantar fasciitis were recruited. Patients were excluded if they had a heel injection in the previous 6 months or were currently using custom foot orthoses at the time of screening. Sixtyâ nine patients completed the final followâ up visit 3 months after receiving the footwear intervention.MethodsTreatment failure was considered a <50% reduction in heel pain at 3 month followâ up. Logistic regression models evaluated the possible association between more than 30 clinical and physical examination findings prospectively assessed at enrollment, and treatment response.ResultsInability to dorsiflex the ankle past â 5° (odds ratio [OR] 3.9, P = .024), nonsevere (â ¤7 on ordinal scale) firstâ step pain (OR 3.8, P = .021), and heel valgus in relaxed stance (OR 4.0, P = .014) each predicted treatment failure in multivariable analysis (receiver operating characteristic area under the curve = .769). Limited ankle dorsiflexion also correlated with greater heel pain severity at initial presentation (r = â 0.312, P = .006).ConclusionsPatients with severe ankle equinus were nearly 4 times more likely to experience a favorable response to treatment centered on home Achilles tendon stretching and supportive therapy. Thus, earlier use of more advanced therapies may be most appropriate in those presenting without severe ankle equinus or without severe first step pain. The findings from our study may not be clinically intuitive because patients with less severe equinus and less severe pain at presentation did worse with conservative care.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/146867/1/pmrj436.pd

Dimensionality of Carbon Nanomaterials Determines the Binding and Dynamics of Amyloidogenic Peptides: Multiscale Theoretical Simulations

Experimental studies have demonstrated that nanoparticles can affect the rate of protein self-assembly, possibly interfering with the development of protein misfolding diseases such as Alzheimer's, Parkinson's and prion disease caused by aggregation and fibril formation of amyloid-prone proteins. We employ classical molecular dynamics simulations and large-scale density functional theory calculations to investigate the effects of nanomaterials on the structure, dynamics and binding of an amyloidogenic peptide apoC-II(60-70). We show that the binding affinity of this peptide to carbonaceous nanomaterials such as C60, nanotubes and graphene decreases with increasing nanoparticle curvature. Strong binding is facilitated by the large contact area available for π-stacking between the aromatic residues of the peptide and the extended surfaces of graphene and the nanotube. The highly curved fullerene surface exhibits reduced efficiency for π-stacking but promotes increased peptide dynamics. We postulate that the increase in conformational dynamics of the amyloid peptide can be unfavorable for the formation of fibril competent structures. In contrast, extended fibril forming peptide conformations are promoted by the nanotube and graphene surfaces which can provide a template for fibril-growth

FishNet: an online database of zebrafish anatomy

Background: Over the last two decades, zebrafish have been established as a genetically versatile model system for investigating many different aspects of vertebrate developmental biology. With the credentials of zebrafish as a developmental model now well recognized, the emerging new opportunity is the wider application of zebrafish biology to aspects of human disease modelling. This rapidly increasing use of zebrafish as a model for human disease has necessarily generated interest in the anatomy of later developmental phases such as the larval, juvenile, and adult stages, during which many of the key aspects of organ morphogenesis and maturation take place. Anatomical resources and references that encompass these stages are non-existent in zebrafish and there is therefore an urgent need to understand how different organ systems and anatomical structures develop throughout the life of the fish. Results: To overcome this deficit we have utilized the technique of optical projection tomography to produce three-dimensional (3D) models of larval fish. In order to view and display these models we have created FishNet http://www.fishnet.org.au, an interactive reference of zebrafish anatomy spanning the range of zebrafish development from 24 h until adulthood. Conclusion: FishNet contains more than 36 000 images of larval zebrafish, with more than 1 500 of these being annotated. The 3D models can be manipulated on screen or virtually sectioned. This resource represents the first complete embryo to adult atlas for any species in 3D

The Place of Scripture in the Trajectories of a Distinct Religious Identity among Ravidassias in Britain: Guru Granth Sahib or Amritbani Guru Ravidass

This article highlights narratives, collected as informant testimonies, relating to trajectories of a distinct religious identity among the Ravidassia community in Britain. Current tensions surround the replacement of the Guru Granth Sahib with the Amritbani Guru Ravidass in Ravidassia places of worship. This is primarily in response to cartographies of the Ravidassia identity as distinct from Sikh identity. The opinions of Ravidassia individuals, from a varied age range, expressed in interviews conducted at various periods during 2010–2012, are considered in relation to dominant discourses emphasising the importance of one hegemonic ‘Ravidassia’ scripture. The interview data highlight three main positions among the followers of Guru Ravidass: (1) Ravidassias seeking a distinct identity but preferring to retain the Guru Granth Sahib in Ravidassia places of worship, (2) Ravidassias demanding a distinct identity by installing the Amritbani Guru Ravidass, (3) Ravidassias wanting to maintain their link with the Panth as Sikhs or as Ravidassi Sikhs